TCF12 Antibody / Transcription factor 12 (FY12003)

Flow Cytometry analysis of 293T cells using anti-TCF12 antibody. Overlay histogram showing 293T cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TCF12 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of TCF12 using anti-HEB/TCF12 antibody. Lane 1: human whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: rat thymus tissue lysates, Lane 6: rat C6 whole cell lysates, Lane 7: mouse thymus tissue lysates, Lane 8: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TCF12 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. Expected size of TCF12 ~73 kDa (canonical isoform). Observed band at ~85 kDa is consistent with validated antibody specifications (~80-85 kDa) and likely reflects expression of a longer splice isoform and/or post-translationally modified form of TCF12.